Research On The Anti-invasion And Metastasis Of Xiaoaiping Injection Alone And When Combined With Oxaliplatin And Endostar In Human Hepatoma Cell

Objective:To investigate the effects and the mechanism of Xiaoaiping(XAP) injection alone or combined with oxaliplatin, human recombinant endostatin (endostar) on human hepatoma cell lines SMMC-7721 in vitro, so as, to provide the theoretical foundation for the combination application of chemotherapy with traditional medicine and antiangiogenic therapy to cure hepatocarcinoma.Methods:Observe differences between drug groups and control group of human hepatoma cell lines SMMC-7721 in vitro when they are affected by different concentration of XAP injection, oxaliplatin, endostar alone and by the combinative use of the drugs above. (1) Observe the proliferation effects of drugs on SMMC-7721 by MTT-assay, and select the appropriate concentration for the next experiments. (2) Cell adhesion experiments:Treat SMMC-7721 cells with different drugs concentrations, and contrast the different adhesive ability of cells. (3) Cell migration experiments:Observe the migration capacity of drugs on SMMC-7721 through the cell scratch test. Futherly, perform the quantitative analysis of different groups' migration capacity by Transwell permeable method. (4) Cell invasion experiments:Analyze the difference of cell invasion capacity between different drug groups with Transwell permeable method to.(5)Detecting the expressions of apoptosis-associated gene proteins:E-cadherin,nm23-H1 and MMP-2 with immunohistochemical stain.Results:(1) XAP injection and oxaliplatin could inhibit the proliferation of SMMC-7721 cell lines, and this inhibition depended on time and dose manner. The apoptosis and proliferation rate was less than 30% when we used XAP injection(10μL/mL,15μL/ml,20μL/mL),oxaliplatin(0.5μg/mL) and endostar(6.0μg/mL) alone or by the combinative 24 hours later. (2) The number of SMMC-7721 cells adhesion to Matrigel, FN was reduced to various degrees when the cells were affected by XAP injection(10μL/mL,15μL/mL,20μL/mL). The adhesion inhibition rate showed increasing trend when the concentration of XAP increased. Oxaliplatin, Endostar single drug did not induce significant adhesion inhibition. But When XAP and oxaliplatin was combined to apply, synergism could be observed. (3) XAP, oxaliplatin single drug group can reduce the SMMC-7721 cells in migration capacity (P<0.05, P<0.01, P<0.01, P<0.01). Endostar single drug did not reduce the ability of cell movement. But When XAP and oxaliplatin was used as drug combination manner, the synergism was very significant. (4) XAP, oxaliplatin single drug group can reduce the SMMC-7721 cells in invasive capacity (P<0.05, P<0.01, P<0.01, P<0.01). Endostar single drug did not reduce the ability of cell invasion. However, combination of XAP and oxaliplatin did. (5) XAP can upregulate E-cadherin, nm23-H1 expression, but reduce the expression of MMP-2. Oxaliplatin (0.5μg/ml)can downregulate the MMP-2 expression of SMMC-7721 cells (P<0.01), however, can not affect the expression of E-cadherin and nm23-H1 (P>0.05). Endostar can stimulate nm23-H1 expression (P<0.05), but can not affect E-cadherin, MMP-2 expression.Conclusion:(1) XAP can significantly inhibit the adhesion, migration and invasion capacity of SMMC-7721 cells. The mechanism may relate to upregulation of the E-cadherin, nm23-H1 expression, and reduceMMP-2 expression. (2) Oxaliplatin (0.5μg/ml) showed no significant inhibitive effect on SMMC-7721 cells adhesion ability, but can reduce the SMMC-7721 cells migration and invasion ability. The mechanism may be associated with reduction of MMP-2. (3) With combination of XAP and oxaliplatin, the inhibition of SMMC-7721 cells'adhesion, migration and invasion ability shows the synergism. The mechanism may be regulated through raising the expression of E-cadherin and reducing the expression of MMP-2.